DNA methyltransferase 3b regulates articular cartilage homeostasis by altering metabolism

Shen, Jie; Wang, Cuicui; Li, Daofeng; Xu, Taotao; Myers, Jason R.; Ashton, John M.; Wang, Ting; Zuscik, Michael J.; McAlinden, Audrey; O’Keefe, Regis J.

doi:10.1172/jci.insight.93612

Cited by 57 publications

(73 citation statements)

References 61 publications

(54 reference statements)

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“…One article reported the use of Agc Cre/Cre mice for their analysis (Maihiot et al, 2015). However, most of the articles do not state if Agc +/Cre or Agc Cre/Cre mice were used for phenotypic analysis (Shen et al, 2017; Liao et al, 2017; Hu et al, 2017; Sinha et al, 2017; Jing et al, 2015, 2016; Liu et al, 2015; Zhou et al 2014; Ono et al, 2014; Jing et al 2014; Henry et al, 2012). Therefore, caution must be exercised when interpreting data using this model as Agc Cre/Cre mice themselves have a skeletal phenotype.…”

Section: Resultsmentioning

confidence: 99%

“…Tamoxifen treatment results in nuclear translocation of the Cre-ERT fusion protein and subsequent excision of a floxed gene. Several studies have utilized the Agc Cre model to investigate the role of target genes in developing cartilage tissue (Shen et al, 2017; Liao et al, 2017; He et al, 2017; Hu et al, 2017; Sinha et al, 2017; Jing et al 2016; Liu et al, 2015; Maihiot et al, 2015; Zhou et al 2014; Ono et al, 2014; Jing et al 2014; 2013; Henry et al, 2012). Prior to breeding with our floxed mice, we noted variable weight and size among the Agc Cre littermates.…”

Section: Introductionmentioning

confidence: 99%

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Dwarfism in homozygous Agc1^CreERT mice is associated with decreased expression of aggrecan

Rashid

Chen

Hassan

et al. 2017

Genesis

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SUMMARY Aggrecan (Acan), a large proteoglycan is abundantly expressed in cartilage tissue. Disruption of Acan gene causes dwarfism and perinatal lethality of homozygous mice. Due to sustained expression of Acan in the growth plate and articular cartilage, AgcCre model has been developed for the regulated ablation of target gene in chondrocytes. In this model, the IRES-CreERT-Neo-pgk transgene is knocked-in the 3′UTR of the Acan gene. We consistently noticed variable weight and size among the AgcCre littermates, prompting us to examine the cause of this phenotype. Wild-type, Cre-heterozygous (Agc+/Cre), and Cre-homozygous (AgcCre/Cre) littermates were indistinguishable at birth. However, by 1-month, AgcCre/Cre mice showed a significant reduction in body weight (18–27%) and body length (19–22%). Low body weight and dwarfism was sustained through adulthood and occurred in both genders. Compared with wild-type and Agc+/Cre littermates, long bones and vertebrae were shorter in AgcCre/Cre mice. Histological analysis of AgcCre/Cre mice revealed a significant reduction in the length of the growth plate and the thickness of articular cartilage. The amount of proteoglycan deposited in the cartilage of AgcCre/Cre mice was nearly half of the WT littermates. Analysis of gene expression indicates impaired differentiation of chondrocyte in hyaline cartilage of AgcCre/Cre mice. Notably, both Acan mRNA and protein was reduced by 50% in AgcCre/Cre mice. A strong correlation was noted between the level of Acan mRNA and the body length. Importantly, Agc+/Cre mice showed no overt skeletal phenotype. Thus to avoid misinterpretation of data, only the Agc+/Cre mice should be used for conditional deletion of a target gene in the cartilage tissue.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Dwarfism in homozygous Agc1^CreERT mice is associated with decreased expression of aggrecan

Rashid

Chen

Hassan

et al. 2017

Genesis

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“…In OA disease, DNMT3B regulates postnatal cartilage homeostasis, and genome‐wide DNA methylation studies have identified epigenomic changes in OA cartilage along with differences between mild and severe OA . Comparisons between young and old human non‐OA and OA samples revealed age‐ and disease‐related epigenetic features; and characterization of the DNA methylome has identified different epigenomic landscapes in hip and knee cartilage and distinct clusters of OA patients .…”

Section: Epigenetic Dysregulation Of Oa Chondrocytesmentioning

confidence: 99%

“…57,102 Since hyper or hypomethylation of DNA can modulate gene transcription, altered DNA methylation is often associated with aberrant cellular phenotypes and abnormal gene expression in several pathologies, 138 including OA. 92 In OA disease, DNMT3B regulates postnatal cartilage homeostasis, 139 and genome-wide DNA methylation studies have identified epigenomic changes in OA cartilage 99,100,102 along with differences between mild and severe OA. 140 Comparisons between young and old human non-OA and OA samples revealed age-and disease-related epigenetic features; 141 and characterization of the DNA methylome has identified different epigenomic landscapes in hip and knee cartilage 142 and distinct clusters of OA patients.…”

Section: Figure 4 Modulation Of Chondrocyte Hypertrophy By Canonicalmentioning

confidence: 99%

Phenotypic instability of chondrocytes in osteoarthritis: on a path to hypertrophy

Singh

Marcu

Goldring

et al. 2018

Annals of the New York Academy of Sciences

113

130

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Articular chondrocytes are quiescent, fully differentiated cells responsible for the homeostasis of adult articular cartilage by maintaining cellular survival functions and the fine-tuned balance between anabolic and catabolic functions. This balance requires phenotypic stability that is lost in osteoarthritis (OA), a disease that affects and involves all joint tissues and especially impacts articular cartilage structural integrity. In OA, articular chondrocytes respond to the accumulation of injurious biochemical and biomechanical insults by shifting toward a degradative and hypertrophy-like state, involving abnormal matrix production and increased aggrecanase and collagenase activities. Hypertrophy is a necessary, transient developmental stage in growth plate chondrocytes that culminates in bone formation; in OA, however, chondrocyte hypertrophy is catastrophic and it is believed to initiate and perpetuate a cascade of events that ultimately result in permanent cartilage damage. Emphasizing changes in DNA methylation status and alterations in NF-κB signaling in OA, this review summarizes the data from the literature highlighting the loss of phenotypic stability and the hypertrophic differentiation of OA chondrocytes as central contributing factors to OA pathogenesis.

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“…Similarly DNA methylation may regulate gene expression during chondrogenesis where COLl0Al induction correlates with its promoter demethylation, and intereference with DNA methylation during chondrogenesis by treatment with 5-aza-C alters gene expression (Zimmermann et al , 2008; El-Serafi et al , 2011). While addition of methylation by DNMT3A has been found to regulate SOX9 expression in limb bud mesenchymal cells, and by DNMT3B to regulate cartilage metabolism and homeostasis (Kumar and Lassar, 2014; Shen etal. , 2017).…”

Section: Introductionmentioning

confidence: 99%

DNA hypomethylation during MSC chondrogenesis occurs predominantly at enhancer regions

Barter

Bui

Cheung

et al. 2019

Preprint

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SummaryRegulation of transcription occurs in a cell type specific manner by epigenetic mechanisms including DNA methylation and histone modifications. Methylation changes during stem cell differentiation may play a key role in lineage specification. We sought to characterise DNA methylation changes during chondrogenesis of mesenchymal stem cells (MSCs) in order to further our understanding of epigenetic regulation in chondrocytes. The consequences of which has potential to improve cartilage generation for tissue engineering purposes and also to provide context for observed methylation changes in cartilage diseases such as osteoarthritis. We identified significant DNA hypomethylation during chondrogenesis including changes at many key cartilage gene loci. Importantly characterisation of significant CpG loci indicated their predominant localisation to enhancer regions. Comparison with adult cartilage and other tissue methylation profiles identified chondrocyte-specific regulatory regions. Taken together we have associated methylation at many CpGs with the chondrocyte phenotype.AbstractRegulation of transcription is determined in a cell type specific manner by epigenetic mechanisms including DNA methylation and histone modifications. Methylation changes during stem cell differentiation may play a role in lineage specification. Multipotent mesenchymal stem cell (MSC) differentiation into chondrocytes not only serves as a model for chondrocyte development but also provides an important source of cartilage for tissue engineering purposes. We sought to characterise DNA methylation changes during chondrogenesis to further understanding of epigenetic regulation but to also provide context for the changes identified during disease.DNA cytosine methylation changes during human MSC differentiation into chondrocytes were measured by Infinium 450K methylation array. Methylation changes at gene loci were contrasted with gene expression changes. Chromatin states of significant methylation loci were interpreted by intersection with chondrogenesis histone modification ChlP-seq data. Chondrogenic and cartilage specific hypomethylation was utilised in order to identify a chondrocyte methylome. Articular cartilage and tissue panel DNA methylation was compared and alterations during osteoarthritis cartilage disease classified.Significant DNA hypomethylation was identified following chondrogenic differentiation of MSCs including changes at many key cartilage gene loci. Highly upregulated genes during chondrogenesis were more likely to exhibit a reduction in DNA methylation. Characterisation of significant CpG loci indicated their predominant localisation in CpG poor regions which importantly are most likely to correspond to enhancer regions. Methylation level at certain CpGs following chondrogenesis corresponds to the level found in adult cartilage.Taken together, considerable demethylation changes to the epigenetic landscape occur during MSC chondrogenesis especially at sites marked by enhancer modifications. Comparison with other tissues, including healthy and OA cartilage, associates CpGs to the chondrocyte phenotype and provides context for changes in disease.

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DNA methyltransferase 3b regulates articular cartilage homeostasis by altering metabolism

Cited by 57 publications

References 61 publications

Dwarfism in homozygous Agc1^CreERT mice is associated with decreased expression of aggrecan

Dwarfism in homozygous Agc1^CreERT mice is associated with decreased expression of aggrecan

Phenotypic instability of chondrocytes in osteoarthritis: on a path to hypertrophy

DNA hypomethylation during MSC chondrogenesis occurs predominantly at enhancer regions

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DNA methyltransferase 3b regulates articular cartilage homeostasis by altering metabolism

Cited by 57 publications

References 61 publications

Dwarfism in homozygous Agc1CreERT mice is associated with decreased expression of aggrecan

Dwarfism in homozygous Agc1CreERT mice is associated with decreased expression of aggrecan

Phenotypic instability of chondrocytes in osteoarthritis: on a path to hypertrophy

DNA hypomethylation during MSC chondrogenesis occurs predominantly at enhancer regions

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Dwarfism in homozygous Agc1^CreERT mice is associated with decreased expression of aggrecan

Dwarfism in homozygous Agc1^CreERT mice is associated with decreased expression of aggrecan